Method and composition for management of anaplasmosis

ABSTRACT

A method and composition for the management of anaplasmosis which comprises administering to cattle a composition containing the compound of the formula   D R A W I N G

United States Patent Barrett Nov. 4, 1975 [54] METHOD AND COMPOSITION FOR 3,321,362 5/1967 Lemin 260/552 MAN G OF ANAPLASMOSIS 3,520,924 7/1970 Winkelmann 260/552 [75] Inventor: Paul Anthony Barrett, London,

England [73] Assignee: Burroughs Wellcome Co., Research Triangle Park, NC.

[22] Filed: Sept. 8, 1972 [21] Appl. No.: 287,337

Related US. Application Data [62] Division of Ser. No. 826,038, March 18, 1969, Pat.

[30] Foreign Application Priority Data Mar. 26, 1968 United Kingdom 14529/68 [52] US. Cl. 424/323 [51] Int. Cl. A61K 31/175 [58] Field of Search 424/323 [56] References Cited UNITED STATES PATENTS 3,265,570 8/1966 Michaels 424/323 Primary Examiner-Sam Rosen Assistant ExaminerDaren M. Stephens Attorney, Agent, or FirmDonald Brown [57] ABSTRACT A method and composition for the management of anaplasmosis which comprises administering to cattle a composition containing the compound of the formula wherein R is a methyl or ethyl group, R is a hydrogen atom or a methyl group, and R and R are different, each being a hydrogen atom, a methyl, or an ethyl group.

41 Claims, No Drawings METHOD AND COMPOSITION .FOR MANAGEMENT OF ANAPLASMOSIS This is a division of application Ser. No. 826,038, filed on Mar. 18, 1969, now U.S. Pat. No. 3,709,935.

This invention relates to anaplasmicidal glyoxal dithiosemicarbazones, and to the treatment of anaplasmosis in cattle.

It has already been disclosed in British Patent Specification No. 1,097,333 that a class of glyoxal dithiosemicarbazones derivatives showed activity against anaplasmosis in cattle. This disease is prevalent in large areas of the Asian, African, American and Australasian continents and certain Southern areas of Europe which have a mediterranean type of climate. The infecting organism, Anaplasma marginale, attacks the red blood cells of the cattle, characteristically causing anaemia, general debility, and fever of the animal, which often prove fatal.

It has now been found that two undisclosed members of this class, namely a-ethoxyethylglyoxal l-(4- methylthiosemicarbazone)-2-thiosemicarbazone and a-ethoxyethylgloxal l-thiosemicarbazone-2-(4'- methylthiosemi-carbazone), show an unexpectedly high degree of activity against anaplasmae, when intravenously administered to cattle. The latter compound also manifests unusually high activity when given subcutaneously and produce a good suppression of anaemia. In addition to these advantages, both compounds are less toxic than the best compound of the previously disclosed group. The closest similarly unsymmetrical homologues, with respect to the thiosemicarbazone residues, namely a-ethoxyethylglyoxal l-thiosemicarbazone-2-4'-ethylthiosemicarbazone, a-methoxyethylglyoxal l -4 '-methylthiosemicarbazone-2-thiosemicarbazone, a-methoxyethylglyoxal l-thiosemicarbazone- 2-4'-methylthiosemicarbazone, methoxymethylglyoxal lthiosernicarbazone2-4-rnethylthiosemicarbaxone, and ethoxymethylglyoxal 1-thiosemicarbazone-2-4- methylthiosemicarbazone, also show advantages in this respect.

The present invention therefore provides compounds of formula (III),

H =N. NH. CS. NH. R

wherein R is a methyl or ethyl group, R is a hydrogen atom or a methyl group, and R and R are different, each being an hydrogen atom, a methyl, or an ethyl group. In particular there are provided the compounds hereinbefore listed, especially a-ethoxyethylglyoxal lthiosemicarbazone-2-4'-methylthiosemicarbazone and a-ethoxyethylglyoxal l-4-methylthiosemicarbazone-2- thiosemicarbazone.

In another aspect the present invention provides formulations containing any of the compounds of formula (III), as hereinbefore defined, or of the above list, in association with a therapeutically acceptable carrier. In a further aspect the invention provides a method of treating anaplasmosis in cattle, comprising the parenteral or oral administration of an effective dosage of these compounds to the infected animal.

The compounds according to the present invention may conveniently be prepared by adaptation of methods known, in actual use or described in the literature,

on the subject. I

The methods include standard techniques for producing unsymmetrical dithiosemicarbazones by reacting the corresponding glyoxal with one equivalent of a thiosemicarbazide and then with one equivalent of another thiosemicarbazide, or produce the thiosemicarbazone chains stagewise from other starting materials. The desired compound must, of course, be isolated from the resulting mixture of symmetrical and various unsymmetrical compounds by methods known for separation of mixture.

It hasalso been found that dialkylacetal mono-thiosemicarbazone of formula (II) can most advantageously be used for the preparation of the desired unsymmetrical dithiosemicarbazones by reacting this derivative with a substituted or unsubstituted thiosemicarbazide different from that used in the preparation of formula (II). The novel starting compound of formula (II).

wherein Z and Z are the same or different, and each is an alkyl group, preferably a lower alkyl group having from 1 to 5 carbon atoms, can conveniently be obtained by reacting the corresponding a-alkoxyalkylglyoxal-l-dialkylacetal with thiosemicarbazide, 4- methyl or 4-ethyl-thiosemicarbazide, preferably in a polar solvent such as ethanol. The resulting a-alkoxyalkylglyoxal 1-dialkylacetal-2-thiosemicarbazone or 2-(4-methylor ethylthiosemicarbazone) is conveniently re acted with 4-methylor 4-ethylthiosemicarbazide or thiosemicarbazide, respectively, in ethanol in the presence of a strong acid. The compounds are sparingly soluble and separate from the reaction mixture. They can be obtained in a pure form by recrystallization, for instance from water containing some alcohol.

The products obtained by these methods are substantially homogeneous, but may be contaminated with small amounts of the two corresponding symmetrical dithiosemicarbazones. If required, these can be removed by special purification methods known in the art. This is, however, hardly necessary, since the con taminents hav similar properties and their effect is insignificant.

The compounds according to the present invention may be presented in the formulations suitable for oral or parenteral administration. For example, the oral preparations may be tablets, capsules, granules, powder, suspension, or solutions, which may contain diluents, binding agents, dispersing agents, surface-active agents, lubricating agents, coating materials, colouring agents, solvents, thickening agents, suspending agents, or other therapeutically acceptable additives, and these preparations may be presented in unit-dose form or multi-dose form, or as additives to feedstuffs. The injectable form may be anon-aqueous solution for instance in dimethyl sulphoxide or dirnethylacetarnide or suspension in a therapeutically acceptable liquid or mixture of liquids, which may contain bacteriostatic agents, antioxidants, buffers, solutes to render the solution isotonic with the blood, thickening agents, suspending agents, or other therapeutically acceptable additives. Such preparations are presented in unit-dose forms such as ampoules or disposable injection devices, or in multi-dose forms such as a bottle from which the appropriate doses may be withdrawn. All such preparations should preferably be rendered sterile.

For injections the compounds may be presented in the form of a sterile suspension containing up to 20% finely dispersed material in an acceptable medium. Preferably the formulation contains 10% dispersed material at an average particle size below 10pm.

Usually the dosage representing about 20 ing/kg. (orally) or 10 mg./kg. (intravenously or subsutaneously) is sufficient for this purpose; significant improvements can occasionally be achieved with considerably lower dosages, such as about mg./kg. or even 2.5 mg./kg. v

Preferably the compounds according to the present invention, particularly oz-ethoxyethylglyoxal l-thiosemicarbazone-2-4'-methylthiosemicarbazone, can be advantageously presented for injection in a solution of dimethyl sulphoxide or dimethylacetamide, to which propylene glycol has been added. For instance, a to 20% solution of the active compound may be produced by dissolving the material in 2 to 8 volume parts of dimethyl sulphoxide or dimethylacetamide and making up the volume with propylene glycol to produce the required concentration.

The natural incubation period for anaplasmosis is about 20 to 50 days in cattle. Thus the smallest animal likely to be treated would be a seven-weeks old calf which would weigh about 40 to 50 kg. or more. Treating such an animal would require 200 to 500 mg. at a dose of about 5 to 10 mg. /kg., but adult beasts would, of course, require a much larger dose.

In a further aspect, the present invention provides a method of preventing anaplasmosis in cattle, comprising the repeated intramuscular, subcutaneous or oral administration of the compounds as hereinbefore defined, in a dosage of abput 1 mg.kg. per day to the animal for at least 10 days. In a further aspect, there is provided a method of sterilising the infection with A. marginale in carrier cattle comprising the oral administration of these compounds at a dosage of about 1 mg./kg. for a longer period.

The following examples illustrate the invention:

EXAMPLE 1 To a-ethoxyethylglyoxal diethylacetal (prepared by the method of Tiffany et al., J. Amer. Chem. Soc., (1957) 79, 1687) (61.2 g.) in absolute ethanol (300 ml.) was added finely ground thiosemicarbazide (27.3 g.) and the mixture stirred at room temperature for 3 days. The thiosemicarbazide had all passed into solution after 48 hours.

To the solution of a-ethoxyethylglyoxal diethylacetal 2-thiosemicarbazone so obtained was added finely ground 4-methylthiosemicarbazide (31.5 g.), and the mixture stirred and cooled in an ice-bath. Concentrated hydrochloric acid (30 ml.) was added dropwise over 30 minutes, and the mixture stirred at 0C for a further 6 hours. The mixture gradually thickened up with the separation of dithiosemicarbazone, which was filtered off, washed with a mixture of ethanol and light petroleum, and dried in vacuo over sodium hydroxide. The crude material (62 g., mp. 190C) was recrystal- 4 lised from water to which a little ethanol was added, to give a-ethoxyethylglyoxal l-(4'-methylthiosemicarbazone)-2-thiosemicarbazone as pale yellow needles (51 g.), m.p. 192l93C.

After a solution of a-ethoxyethylglyoxal diethylacetal 2-thiosemicarbazone prepared as described in the first paragraph is evaporated to dryness in vacuo an oil remains which solidifies on stirring with petrol, and may be recrystallised from ether/petrol to give the compound as colourless crystalls, m.p. 68C.

EXAMPLE 2 A mixture of a-ethoxyethylglyoxal diethylacetal (61.2 g.) and finely ground 4-methylthiosemicarbazide (31.5 g.) in absolute ethanol (100 ml.) was stirred at room temperature for 3 days. The 4-methylthiosemicarbazide was passed rapidly into solution. To the solution so obtained was added finely ground thiosemicarbazide (27.3 g.) and the mixture was stirred and cooled at 0C while concentrated hydrochloric acid (30 ml.) was added dropwise over 15 minutes and stirred at 0 for a further 6 hours. The precipitated dithiosemicarbazone was filtered off, washed with a mixture of ethanol and light petroleum and recrystallised from a mixture of equal parts of ethanol and water to give a-ethoxyethylglyoxal 1-thiosemicarbazone-2-(4'- methylthiosemicarbazone) as a yellow crystalline powder (35 g.), m.p. 186-7C (decomp.).

Alternatively, after stirring for 6 hours, the hydrochloric acid may be neutralised by the addition of saturated aqueous carbonate solution until the mixture is just alkaline to Billiant Yellow paper. The precipitated solid is then filtered off, washed with 50% aqueous ethanol and with water until free of inorganic material, dried in vacuo, and recrystallised to give the compound as described above.

EXAMPLE 3 a-Ethoxyethylglyoxal l-thiosemicarbazone-2-4'- methyl-thiosemicarbazone was also prepared, using aethoxyethylglyoxal dimethylacetal (52.8 g.) (prepared by the method of Tiffany et al. (loc. cit.) using methanol instead of ethanol, and having b.p. 4546C/0.05 mmHg) in place of the diethylacetal used in Example 2.

EXAIVIPLE 4 By the method of Example 2, using 4-ethylthiosemicarbazide in place of 4-methylthiosemicarbazide, was obtained a-ethoxyethylglyoxal 1-thiosemicarbazone 2-4'-ethylthiosemicarbazone as pale yellow crystals having m.p. ll 76C(decomp.)

EXAMPLE 5 a-Methoxyethylglyoxal diethylacetal (prepared from a-methoxyethylglyoxal by the methodzof Tiffany et al. (loc. cit.) and having b.p. l00l05C-/14 mml-lg) was reacted successively with equimolecular amounts of thiosemicarbazide and 4-methylthiosemicarbazide by the method of Example 1 to give a-methoxyethylglyoxal l-4-methyl thiosemicarbazone-2-thiosemicarbazone as pale yellow crystals, mp. 186C (decomp.).

EXAMPLE 6 a-Methoxyethylglyoxal diethylacetal, described in Example 5, was reacted successively with equimolecular amounts of 4-methyl-thiosemicarbazide and thiosemicarbazide by the method of Example 2 to give amethoxyethylglyoxal l-thiosemicarbazone-2-4'- methylthiosemicarbazone as pale yellow crystals having mp. 205C (decomp).

EXAMPLE 7 EXAMPLE 8 Ethoxymethylglyoxal diethylacetal (prepared by the method of Tifi'any et al. (loc. cit.) from ethoxymethylglyoxal, and having the b.p. 104l08C/12 mmHg) was reacted successively with equimolecular amounts of 4-methylthiosemicarbazide and thiosemicarbazide by the method of Example 2 to give ethoxymethylglyoxal 1-thiosemicarbazone-2-4'-methylthiosemicarbazone as pale yellow crystals having mp. 180C (decomp.)

EXAMPLE 9 A solution for injection was prepared by using the following materials:

a-ethoxyethylglyoxal l-thiosemicarbazone-2-4' methyl-thiosemicarbazone 10.0 g Dimethyl Sulphoxide (analytical reagent guide) 50.0 ml Propylene Glycol B.P. to make 100 ml.

The active ingredient was completely dissolved in di methyl sulphoxide by stirring, and the volume was made up to 100 w. with propylene glycol. The solution was sterilized by filtration through a solvinert millipove membrane filter under aseptic conditions.

The sterile solution was aseptically distributed into sterile 50 ml-vials. The vials were closed with sterile transparent latex plugs.

EXAMPLE 10 A solution for injection was prepared according to the method described in Example 9 by using the following formulation:

EXAMPLE 11 A solution for injection was prepared according to the method described in Example 9 by using the following formulation:

a-ethoxyethylglyoxal l-thiosemicarbazone- -2-4'-methylthiosemicarbazone 20.0 g.

Dimethyl Acetamide (analytical reagent grade) 40.0 ml.

Propylene Glycol B.P. to make 100 ml.

EXAMPLE 12 Tablets of a-ethoxyethylglyoxal l-thiosemicarbazone-2-4-methylthiosemicarbazone were made by granulating the compound (0.5 g.) in a fine powder with gelatin in alcohol and water, adding magnesium stearate (0.005 g.) as a lubricant, and compressing the mixture directly.

Example 13 Tablets of a-ethoxyethylglyoxal l-thiosemicarbazone-2-4'-methylthiosemicarbazone were made by mixing the compound with a diluent (lactose), a dispersing agent (starch), and a surface active agent (polyoxyethylene sorbitan monolaurate). The mixture was granulated with a 5.0% starch mucilage, dried, mixed with a lubricating agent (magnesium stearate) and compressed a-ethoxyethylglyoxal l-thiosemicarbazone -2-4'-methylthiosemicarbazone Lactose Starch Poloxyethylene sorbitan monolaurate Magnesium stearate EXAMPLE 14 Three splenectomised oalves were intravenously infected with 5 ml. blood parasitized with A. marginale. The proportion of erythracites was 24 to 37% in the donor animals. Five days later, the infected calves were intravenously administered l0. rng./kg. of a a-ethoxyethylglyox al 1 4 '-m ethyl-thiosemicarbazone )-2-thiosemicarbazone. I L

Results were as follows:

Animal Dose Route Eryth. lnfecd Hb g/lOOml Death No. D Day D Max. Day D Min.

2810 21 25 H2 l0.l

a-ethoxyethylglyoxal l-thiosemicarbazone- -2-4'-methylthiosemicarbazone 10.0 g. Dimethyl Sulphoxide (analytical reagent grade) 80.0 ml Propylene Glycol B.P. to make 100.0 ml

Very high degree of activity was observed with minimal fall in Hb.

EXAMPLE 15 Three splenectomised calves were intravenously infected with 5 ml. blood parasitized with A. marginale. The proportion of erythracites was 21 to 24% in the donor animals. Five to six days later, the infected calves were subcutaneously injected with 10 mg./kg. of aethoxyethylglyoxal l-thiosemicarbazone-2-(4'-methylthiosemic arbazone Results were as follows:

Animal Dose Route Eryth. lnfec'd Hb g/IOOml Death No. Day D Max. Day D Min.

2824 a/c 2.3 9 l 1.2 9.8 2807 3.2 4.5 12.2 10.9 2819 22 10.5 6.7

A high degree of activity and a good suppression of anaemia was observed.

A high degree of activity against anaplasms with good suppression of anaemia was observed. There was no ev- 1dence of drug persisting at the injection site on autopsy EXAMPLE 16 in 3185 and 3184. Three splenectomised calves were intravenously infected with 5 ml. blood parasitized with A. marginale. The proportion of erythracites was 18% in the donor l5 EXAMPLE 19 animals. Five to seven days later, the infected calves Three splenectomised calves were infected, as in Exwere intravenously injected with 10 mg./kg. of a-ethoxample l6, and were subsequently intravenously inyethylglyoxal l-thiosemicarbazone-2-(4'-methyl-thijected with a-methoxyethylglyoxal-l-thiosemicarbaosemicarbazone). zone-2-4-methylthiosemicarbazone.

Results were as follows: Results were as follows:

Animal Dose Route Eryth. lnfecd Hb/lOOml Death No. D Day D Max. Day D Min.

Animal Dose Route Eryth. lnfec'd Hb g/ 100ml Death No. D Day D Max. Day D Min.

A very high degree of activity and satisfactory sup- High degree of activity against anaplasms was obpression of anaemia were observed. Serve i EXAMPLE l7 Again, three splenectomised calves were intrave- What 1 Claim is; v nously infected with 5 ml. blood parasitized with A. 40 pharmaceutical composition 1n u n1t dosage form marginale, to provide controls for the experiments de- Suitable for Oral par nteral admlmstratlon 1n the scribed in Examples 3 to 5. The proportion of erythrage e t Of anaplasmosls in cattle containing an Cites was 13 to 24% i h donor i l effective anaplasmosis treatment, preventlon or steril- The results were as follows: ization amount of a compound of the formula Animal Dose Route Eryth lnfecd Hb g/lOOml Death No. Day D Max. Day D Min.

2809 Untreated 78 3.2 2808 92 4.2 2826 95 3.6

All three control animals died. 2

l RO-CH- N.N. H.cs.N1-|.R= EXAMPLE 18 1-1 =N.NH.CS.NH.R Three splenectomised calves were infected, as in Example 16, and were subsequently intravenously inwherein R is a methyl or ethyl group, R is a hydrogen jected with a-methaxyethylglyoxal-l-4-methylthiatom or a methyl group, and R and R are different, osemicarbazone-2-thiosemicarbazone in propylene glyeach being a hydrogen atom, a methyl, or an ethyl col/dimethyl sulphoxide solution. group in association with a pharmaceutically accept- Results were as follows: able carrier therefor.

Animal Dose Route Eryth. lnfec'd Hb g/lOOml Death No. D Day D Max. Day D Min.

2. A composition according to claim; 1 in which the compound is a-Ethoxyethylglokal' "l thios'emicarba zone-2-4-methylthiosemicarbazone.';; v

3. A composition according to claim 1 ..in';.which.;the ii compound is a-ethoxyethylglyoxal 1 4. osemicarbazone-2 thio'semicarbazonea 4. A composition for injection accord'mgto 1,, wherein thecarrier is dimethyl sulphoxide orj dimethyl acetamide. I v 1 g f 5. A composition according to claim-4, which con; tains up to to 60% w/v propylene glycol.

6. A composition according to claim 4, wherein the resulting concentration of the compound is about 10 to w/v.

7. A formulation according to claim 1, containing a unit dose of about 200 to 500 mg. of the compound.

8. A method for the treatmnet of anaplasmosis in cattle, comprising the oral or parenteral administration of an effective anaplasmosis treatment dosage of a compound of the formula wherein R is a methyl or ethyl group, R is a hydrogen atom or a methyl group, and R and R are different, each being a hydrogen atom, a methyl, or an ethyl group to the infected animal.

9. A method for preventing anaplasmosis in cattle comprising the repeated intramuscular, subcutaneous or oral administration of an effective anaplasmosis prevention dosage of a compound of the formula wherein R is a methyl or ethyl group, R is a hydrogen atom or aa methyl group, and R and R are different, each being a hydrogen atom, a methyl, or an ethyl group to the animal.

10. A method of sterilising carrier cattle infected with A. marginale of the A. marginale microorganism comprising the oral administration of the compound of the formula 10 p 13. A rnethod according tovclaim 8 wherein the compound is a-Ethoigyethylglyoxal l-4-methylthiosemicarba z orie-2-thiosemicarbazone. 14. A method according to claim 8 wherein the compoundjs az-Ethoxyethyl'glyoxal l-thiosemicarbazone-2- 4Qethylthiosemicarbazone.

15. A method according to claim 8 wherein the com- 1-4 '-methylthipound is a-Methoxyethylglyoxal osemicarbazone-2- thiosemicarbazone.

16'.- A method according to claim 8 wherein the compound is a-Methoxyethylglyoxal l-thiosemicarbazone- 2-4'methyl-thiosemicarbazone.

17. A method according to claim 8 wherein the compound is a-Methoxymethylglyoxal l-thiosemicarbazone-2-4'methyl-thiosemicarbazone.

18. A method according to claim 8 wherein the com pound is a-Ethoxymethylglyoxal l-thiosemicarbazone- 2-4-methyl-thiosemicarbazone.

19. A method according to claim 9 wherein the compound is a-Ethoxyethylgloxal 1-thiosemicarbazone-2- 4-methylthiosemicarbazone.

20. A method according to claim 9 wherein the compound is a-Ethoxyethylglyoxal 1-4'-methylthiosemicarbazone-2thiosemicarbazone.

21. A method according to claim 9 wherein the compound is a-Ethoxyethylglyoxal l-thiosemicarbazone-Z- 4'-ethylthiosemicarbazone.

22. A method according to claim 9 wherein the compound is a-Methoxyethylglyoxa] l-4-methylthiosemicarbazone-Z-thiosemicarbazone.

23. A method according to claim 9 wherein the compound is a-Methoxyethylglyoxal l-thiosemicarbazone- 2-4-methylthiosemicarbazone.

24. A method according to claim 9 wherein the compound is a-Methoxymethylglyoxal l-thiosemicarbazone-2-4'methyl-thiosemicarbazone.

25. A method according to claim 9 wherein the compound is a-Ethoxymethylglyoxal l-thiosemicarbazone- 2-4'-methyl-thiosemicarbazone.

26. A method according to claim 10 wherein the compound is a-Ethoxyethylgloxal l-thiosemicarbazone-2-4'methylthiosemicarbazone.

27. A method according to claim 10 wherein the compound is a-Ethoxyethylglyoxal 1-4-methylthiosemicarbazone-2-thiosemicarbazone.

28. A method according to claim 10 wherein the compound is a-Ethoxyethylglyoxa] l-thiosemicarbazone-2-4-ethylthiosemicarbazone.

29. A method according to claim 10 wherein the compound is bz-Methoxyethylglyoxal l-4'-methylthiosemicarbazone- 2-thiosemicarbazone.

30. A method according to claim 10 wherein the compound is a-Methoxyethylglyoxal l-thiosemicarbazone-2-4'methyl-thiosemicarbazone.

31. A method according to claim 10 wherein the compound is a-Methoxymethylglyoxal l-thiosemicarbaz one-2-4-methyl-thiosemicarbazone.

32. A method according to claim 10 wherein the compound is a-Ethoxymethylgloxal l-thiosemicarbazone-2-4'-methylthiosemicarbazone.

33. A composition according to claim 1 in which the compound is a-Ethoxyethylglyoxal l-4-methylthiosemicarbazone 2-thiosemicarbazone.

34. A composition according to claim 1 in which the compound is a-Methoxyethylglyoxal l-4'-methylthiosemicarbazone-Z-thiosemicarbazone.

35. A composition according to claim 1 in which the compound is a-Methoxyethylglyoxal l-thiosemicarbazone- 2-4 '-methyl-thiosemicarbazone.

36. A composition according to claim 1 in which the compound is a-Methoxymethylglyoxal l-thiosemicap' bazone-2-4'-methyl-thiosemicarbazone.

37. A composition according to claim 1 in which the compound is a-Ethoxymethylglyoxal l-thiosemicarbazone-2-4'-methyl-thiosemicarbazone.

38. A composition according to claim 2 wherein the effective amount is an effective anaplasmosls treatment amount. u v

39. A composition according to claim 2 wherein the effective amount is an effective anaplasrnosis prevenzone-2-4'-ethylthiosemicarbazone. 

1. A PHARMACEUTICAL COMPOSITION IN UNIT DOSAGE FORM SUITABLE FOR ORAL OR PARENTERAL ADMINISTRATION IN THE MANAGEMENT OF ANAPLASMOSIS IN CATTLE CONTAINING AN EFFECTIVE ANAPLASMOSIS TREATMENT, PREVENTION OR STERILIZATION AMOUNT OF A COMPOUND OF THE FORMULA
 2. A composition according to claim 1 in which the compound is Alpha -Ethoxyethylgloxal 1-thiosemicarbazone-2-4''-methylthiosemicarbazone.
 3. A composition according to claim 1 in which the compound is Alpha -ethoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 4. A composition for injection according to claim 1, wherein the carrier is dimethyl sulphoxide or dimethylacetamide.
 5. A composition according to claim 4, which contains up to 10 to 60% w/v propylene glycol.
 6. A composition according to claim 4, wherein the resulting concentration of the compound is about 10 to 20% w/v.
 7. A formulation according to claim 1, containing a unit dose of about 200 to 500 mg. of the compound.
 8. A method for the treatmnet of anaplasmosis in cattle, comprising the oral or parenteral administration of an effective anaplasmosis treatment dosage of a compound of the formula
 9. A method for preventing anaplasmosis in cattle comprising the repeated intramuscular, subcutaneous or oral administration of an effective anaplasmosis prevention dosage of a compound of the formula
 10. A METHOD OF STERILISING CARRIER CATTLE INFECTED WITH A. MARGINALE OF THE A. MARGINALE MICROORGANISM COMPRISING THE ORAL ADMINISTRATION OF THE COMPOUND OF THE FORMULA
 11. A method according to claim 8, comprising the oral administration of about 20 mg/kg. to the infected animal.
 12. A method according to claim 8 wherein the compound is Alpha -Ethoxyethylgloxal 1-thiosemicarbazone-2-4''-methylthiosemicarabazone.
 13. A method according to claim 8 wherein the compound is Alpha -Ethoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 14. A method according to claim 8 wherein the compound is Alpha -Ethoxyethylglyoxal 1-thiosemicarbazone-2-4''-ethylthiosemicarbazone.
 15. A method according to claim 8 wherein the compound is Alpha -Methoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 16. A method according to claim 8 wherein the compound is Alpha -Methoxyethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 17. A method according to claim 8 wherein the compound is Alpha -Methoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 18. A method according to claim 8 wherein the compound is Alpha -Ethoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 19. A method according to claim 9 wherein the compound is Alpha -Ethoxyethylgloxal 1-thiosemicarbazone-2-4''-methylthiosemicarbazone.
 20. A method according to claim 9 wherein the compound is Alpha -Ethoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 21. A method according to claim 9 wherein the compound is Alpha -Ethoxyethylglyoxal 1-thiosemicarbazone-2-4''-ethylthiosemicarbazone.
 22. A method according to claim 9 wherein the compound is Alpha -Methoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 23. A method according to claim 9 wherein the compound is Alpha -Methoxyethylglyoxal 1-thiosemicarbazone-2-4''-methylthiosemicarbazone.
 24. A method according to claim 9 wherein the compound is Alpha -Methoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 25. A method according to claim 9 wherein the compound is Alpha -Ethoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 26. A method according to claim 10 wherein the compound is Alpha -Ethoxyethylgloxal 1-thiosemicarbazone-2-4''-methylthiosemicarbazone.
 27. A method according to claim 10 wherein the compound is Alpha -Ethoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 28. A method according to claim 10 wherein the compound is Alpha -Ethoxyethylglyoxal 1-thiosemicarbazone-2-4''-ethylthiosemicarbazone.
 29. A method according to claim 10 wherein the compound is Alpha -Methoxyethylglyoxal 1-4''-methylthiosemicarbazone-2-thiosemicarbazone.
 30. A method according to claim 10 wherein the compound is Alpha -Methoxyethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 31. A method according to claim 10 wherein the compound is Alpha -Methoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 32. A method according to claim 10 wherein the compound is Alpha -Ethoxymethylgloxal 1-thiosemicarbazone-2-4''-methylthiosemicarbazone.
 33. A composition according to claim 1 in which the compound is Alpha -Ethoxyethylglyoxal 1-4''-methylthiosemicarbazone -2-thiosemicarbazone.
 34. A composition according to claim 1 in which the compound is Alpha -Methoxyethylglyoxal 1-4''-methylthio-semicarbazone-2-thiosemicarbazone.
 35. A composition according to claim 1 in which the compound is Alpha -Methoxyethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 36. A composition according to claim 1 in which the compound is Alpha -Methoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 37. A composition according to claim 1 in which the compound is Alpha -Ethoxymethylglyoxal 1-thiosemicarbazone-2-4''-methyl-thiosemicarbazone.
 38. A composition according to claim 2 wherein the effective amount is an effective anaplasmosis treatment amount.
 39. A composition according to claim 2 wherein the effective amount is an effective anaplasmosis prevention amount.
 40. A composition according to claim 2 wherein the effective amount is an effective anaplasmosis sterilisation amount.
 41. A composition according to claim 1 in which the compound is Alpha -Ethoxyethylglyoxal 1-thiosemicarbazone-2-4''-ethylthiosemicarbazone. 